[CLASS XII- BIOLOGY] 2024-2025

Experiment No -1

(2) Materials required: Flower, needles, Safranine stain, glycerine, coverslips,

microscope.

(3) Procedure:
(1) Take out the pistil from a Portulaca flower.
(2) Stain the isolated pistil and mount in glycerine, Press gently, observe under dissecting
microscope.
(3) Take out carefully each germinated pollen and count.
(4) Mount a pollen in glycerine and observe under compound microscope.

(4) Observations:
(1) Observe many pollen grains germinating over stigma. The growth of the pollen tube is
stimulated by sugary substances secreted by the stigma.
(2) Pollen tube carrying with it tube nucleus and the generative nucleus.
(3) The generative nucleus divides forming two male gametes.
(4) Count the number of germinated pollen grains.
(5) Precautions:
(i) Mounting should be free from air bubbles.
(ii) Material should be moderately stained.
(iii) Use the clean slide.

Experiment No- 2

(1) Aim: To determine the population density of plants at a place by the quadrate method.

(2) Materials required: Meter scale, string, nails" hammer, measuring, tape, paper.

(3) Principle: Average number of particular plant species present per unit area is called as population
density.

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(4) Method:
(1) Choose a nearly area of square field of size 2m and fix the iron Nails at the comers of this square field.
Now tie a string and ready the square field.
(2) Now divide this square field into 10 small squares by tying strings at the distance of 10- 10
ems. This square is called as quadrate.
(3) Record the name and number of all species present in the squares.
(4) Population density of the plants in this quadrats can be identified by following formulae-
Total no. plant speices
Density =
Observation table:
Observation table
S.No. Name of Plants No.of plants in quadrate of 1x1 m Total no. of Average
species plants in a
quadirate
1 Plant A Quadrate I Quadrate II Quadrate III
2 Plant B
3 Plant C
4 Plant D
Result:
(1) No. of plant species studied in a quadrate are '
(2) Plant species with high density in the quadrate are and species with less density are _.

Precautions :
(1) Only the individuals of one plant species should be considered at one time.
(2) Square field should be taken from single place only.

Experiment No - 3
1. Aim: To make a temporary mount of the onion root tip to study various stages of mitosis. 2.
Materials required: Onion root tips, needles, brush, slide, coverslip, burner, microscope,
acetocarmine stain.
3. Procedure:
(1) Take a root tip on clean slide and put a drop of acetocarmine stain on it.
(2) This makes the stain specific for nuclear materials. Gently warm it a little over a burner. On warming
the stain evaporates. Before it is dried add.more stain on it.
(3) Squash the root tip with the help of a needle or a force put the coverslip. Tap it a bit more from above.
(4) Now take the slide in the folds of a blotting paper and apply gentle pressure with hands.
(5) Observe the slide under the microscope first in the low power and then after locating a specific area in
the high power. Examine and identify various stages of mitosis.

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4. Precautions:

(1) The slide should be warmed gently much above the flame of the spirit lamp.
(2) The acetocarmine stain should be filtered before use.
(3) There should be no air bubble in the slide.

5. Observation:
Various stages of mitosis could be seen –

(1) Interphase:
(i) Chromatin fibres appear in the form of a network within the nucleus. (ii)
Nuclear envelope and nucleolus are distinct.

(2) Prophase:
(1) Chromatin material shortens and condenses into thread like structures called chromosomes. .
(2) Each chromosome consists of two chromatids that are joined at a point called centromere.
(3) Nuclear membrane and nucleolus disappear.

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 3) Metaphase:
(1) Chromosomes become arranged at the equator of the spindle.
(2) Each chromosome get attached to the spindle fibres at its centromere.
(4) Anaphase:
(1) The two sister chromatids of each chromosome separate from the centromere and move towards the
opposite poles.
(2) The daughter chromosomes appear V,J,L and I shaped depending upon the position of centromere.

(5) Telophase:
(1) Nuclear membrane and nucleolus reappears and two daughter nuclei appear at opposite poles.
(2) Cytokines is occurs by cell plate formation between the two daughter nuclei.

Experiment No -4
Isolation of DNA from available plant material such as spinach, green pea seeds, papaya etc.

Spinach leaves/Pea seeds/Papaya, Sand ,test tube, 50 ml beakers, Cheesecloth, Mortar and pestle,
1Omlgraduated cylinder.

95% Ethanol solution (keep ice cold in plastic bottle in freezer),12% Salt solution,29.2 g deionized
salt,250 ml distilled water,50% Detergent solution,50 ml Wisk Free,50 ml distilled water, Contact Lens
Cleaning Solution, Use 1 tablet per 3ml of distilled water.
1. Choose 2-3 spinach leaves. Remove any stems if present.

2. Place 1 ml of distilled water in a mortar and pestle along with leaves. Add a small pinch of sand and
grind until spinach looks like creamed spinach. Add the contents of the mortar and pestle to a 50 ml
beaker.
3. Add 1 ml of 50% detergent solution and 9 ml salt solution to spinach. Mix well with a glass stir rod.
4. Place on a hot plate and heat until boiling. Remove from heat and let sit for minutes.

5. Put on ice for 5 minutes so that it cools down.

6. Pour spinach mixture (supernatant) through cheesecloth into a clean beaker.

7. Pour the supernatant into a test tube then add 1 ml of freshly prepared contact lens cleaning solution.
8. Carefully layer 6 ml chilled 95% ethanol solution onto the green supernatant using a 10 ml graduated
cylinder. Slowly pour ethanol down the side of the test tube. Try not to let the two layers mix together.
9. Using the wire loop, spool the DNA by gently swirling the loop at the interface between the green
supernatant and the clear ethanol. The DNA will congeal at the point where the two layers meet.

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 Spotting 1
Identification: Pollen germination on a slide.

Comments:
(1) Pollen grain or microspore is the first cell of male gametophyte.
(2) Each pollen grain of a flowering plant (angiosperm) possesses two cells.
(i) Vegetative cell (2) Generative cell
(3) On the stigma, the pollen grain absorbs water and nutrients from the stigmatic secretion through its germ
pores.
(4) The tube cell gives rise to a pollen tube, the generative cell also descends into the pollen tube and divides
in to two male gametes.
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 (5) There is only one pollen tube from one pollen.
(6) Certain pollen grain do not germinate and are referred as sterile pollens.

Spotting 2

Identification: T.S of Blastula through permanent slides .

Comments:
(1) It is a spherical mass of about 32 or 64 cells.
(2) It is composed of an outer envelope of cells, the trophoblast or trophoectoderm and inner cell mass
(embryoblast).
(3) Within the envelope there is a fluid filled cavity called blastocoel.
(4) The side of the blastocyst to which the inner cell mass is attached is called the embryonic or animal
pole, while the opposite side is the abembryonic pole.
(5) The inner cell mass is the precursor of the embryo.
(6) In this state it forms the connection with mother's uterus wall which is called implantation.

Common disease causing organisms

Spotting 3 (a)
Identification: Entamoeba
Comments:
(l) It is a human parasite that resides in the upper part of the large Intestine.
(2) It causes the disease called amoebic dysentery or amoebias is.
(3) The symptoms of the diseases Include abdominal pain, repeated motions with blood and mucus.
(4) The parasite is unicellular and has a blunt pseudopodium.
(5) There is a single nucleus and a number of food vacuoles.
(6) It feeds on red blood corpuscles by damaging the wall of large intestine and reaching the blood capillaries.
(7) It produces ulcers and can also reach other body organs.

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 Spotting 3 (b)
Identification : Plasmodium vivex (Malarial parasite)

Comments:
(I) It is a protozoan digenic endoparasite of man.
(2) Its primary host is man and female anopheles is its secondary host.
(3) Plasmodium enters human body in sporozoite stage by the bites of female anopheles.
(4) The sporozoite is spindle shaped and uninucleate organism capable of wriggling movement.
(5) The sporozoites infect liver cell and produce meta-cryptomerozoites.
(6) The metacryptomerozoites enter RBCs, and passes trophozoite signet ring stage and amoeboid
stage and produce schizont and merozoites.
(7) The merozoites enter fresh RBCs and produce gametocytes.

Spotting 3 (c)
Identification: Ascaris
Comments:
(1) It is an endoparasite of the small Intestine of human beings and is more common in childre
(2) The animal shows sexual dimorphism with separate male and female individuals.
(3) The life history is simple and without any intermediate host. The infection occurs through contaminated
food and water.
(4) Ascaris causes abdoininal discomfort and colic pain.
(5) The patient may also suffer from impaired digestion, diarrhoea and vomiting. (6) In children mental
efficiency is affected and body growth is retarded.
(7) It causes ascariasis.

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 Spotting 3 (d)
Identification- Ringworm Pathogen
- Trichophyton sp.
Disease - Athelete foot

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Symptoms:
1. It forms lesions on hairy parts of smooth skin.
2. It also infects the nails of the hands and feet.
3. Some species of these fungi cause ringworm of the scalp found chiefly in children.
4. Mostly they infect the skin so this fungi and disease are called dermatomycoses.
5. Skin becomes dry and whitish in colour with keratin substances.

Spotting 4
Objective:-

To study symbiotic association in root nodules of leguminous plant,cuscuta on host and Lichens through model
/ specimen/ chart

Requirements:-

Model / specimen / chart showing symbiotic association in root nodules of leguminous plant (pea plant),
cuscuta on host and a lichen.

Theory

In nature animals, plants and microbes do not and cannot live in isolation but interact in various ways.

1. When two different species interact with each other (Interspecific interaction) and are in close
association. With each other in such a way that at least one species is benefited while for other
species the relationship may be positive, negative or neutral such association is called symbiotic
relationship

2. There are three basic types of symbiotic relationship

Mutualism (both are benefited)
Commensalism (one species is benefited while other is neither benefited nor harmed) Parasitism
(one species is benefited and other is harmed)

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Observations

1.Rhizobium in root nodules of leguminous plant ( pea plant)

i. Rhizobium bacteria are present in root nodules of leguminous plant and form a
symbiotic relationship,mutualism, where both are benefited from each other.Nitrogen
fixing bacteria Ike Rhizobium fixes atmospheric nitrogen into nitrogenous compounds
ii. Rhizobium can convert atmospheric nitrogen to ammonia that can be used by pea plant
for growth and development

iii. Bacteria receive nutrients and suitable place to grow from plant.

2.Cuscuta with Host

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 i. Cuscuta commonly called dodder or amerbel and live as stem ectoparasite on
other plants
ii. Cuscuta has no fully expanded form of leaves (scale like leaves are present) and
has no chlorophyll
iii. Stem of cuscuta is thin and slender shaped And It winds around the stem of

host plant.Stem of cuscuta fixes itself to the stem of host plant with special

structures called

iv. Haustoria forms direct connection to the vascular bundles of the host and
withdraw water,carbohydrates and other solutes.

v. Roots of cuscuta are temporary and die as soon as it makes connection with host
plant

vi. Cuscuta can weaken or kill plant and reduce crop yield

3. Lichens

i) Lichens are composite organisms representing a symbiotic association (mutualism) between fungus
and algae
ii) The algal component is known as phycobiont and fungal component is known as mycobiont.

iii) .Algae prepare food for fungi and fungi provide shelter and absorb mineral,nutrients and water for its
partner.

iv) They grow on lands,rocks,tree trunks and walls of houses,like dry vegetation.

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